CN102017896A - Method for quickly breeding Ilex dabieshanensis K Yao.et M.P.Deng in vitro - Google Patents
Method for quickly breeding Ilex dabieshanensis K Yao.et M.P.Deng in vitro Download PDFInfo
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- 241001110383 Ilex dabieshanensis Species 0.000 title abstract description 7
- 238000000338 in vitro Methods 0.000 title abstract description 4
- 238000009395 breeding Methods 0.000 title abstract description 3
- 230000001488 breeding effect Effects 0.000 title abstract description 3
- 230000001954 sterilising effect Effects 0.000 claims abstract description 16
- 230000001939 inductive effect Effects 0.000 claims abstract description 12
- 238000005286 illumination Methods 0.000 claims abstract description 9
- 241000209035 Ilex Species 0.000 claims description 36
- 235000003325 Ilex Nutrition 0.000 claims description 35
- 239000002609 medium Substances 0.000 claims description 20
- 238000004659 sterilization and disinfection Methods 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
- 238000011010 flushing procedure Methods 0.000 claims description 10
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- 229920001817 Agar Polymers 0.000 claims description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 5
- 229930006000 Sucrose Natural products 0.000 claims description 5
- 239000008272 agar Substances 0.000 claims description 5
- 229960002523 mercuric chloride Drugs 0.000 claims description 5
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 5
- 238000010899 nucleation Methods 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000008223 sterile water Substances 0.000 claims description 5
- 239000012879 subculture medium Substances 0.000 claims description 5
- 239000005720 sucrose Substances 0.000 claims description 5
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 claims description 5
- 229940033663 thimerosal Drugs 0.000 claims description 5
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- 238000012546 transfer Methods 0.000 claims description 4
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- 231100000614 poison Toxicity 0.000 claims description 2
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- 241000196324 Embryophyta Species 0.000 description 8
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- 241000209034 Aquifoliaceae Species 0.000 description 2
- 235000011848 Ilex opaca Nutrition 0.000 description 2
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- 235000013399 edible fruits Nutrition 0.000 description 2
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- 241001299553 Ilex chinensis Species 0.000 description 1
- 235000003366 Ilex purpurea Nutrition 0.000 description 1
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- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 1
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Abstract
The invention provides a method for breeding Ilex dabieshanensis K Yao.et M.P.Deng in vitro through tissue culture, comprising the following steps of sterilizing an explant, inoculating the sterilized explant to an inducing culture medium for differentiation inducing of buds; controlling the culture temperature to be 20-26DEG C; culturing in a dark environment for 20-26h and then normally culturing at the illumination time of 10-16h/day and the illumination strength of 1,600-2,000Ix for 25-35days; cutting tender branch tips of the Ilex dabieshanensis K Yao.et M.P.Deng and grafting the tender branch tips into a secondary culture medium for carrying out the secondary culture for 25-35days; and connecting aseptic seedlings of Ilex dabieshanensis K Yao.et M.P.Deng into the culture medium for rooting culture at the culture temperature of 20-26DEG C, the illuminaiton time of 10-16h/day and the illumination intensity of 1,600-2.000Ix. By using the method, the problem that the Ilex dabieshanensis K Yao.et M.P.Deng is quickly popularized can be effectively solved.
Description
Technical field
The present invention relates to Dabie Mountain Chinese ilex 1# (Ilex dabieshanensis K Yao.et M.P.Deng.) tissue culture propagation, belong to the technical field of woody plant sapling multiplication.
Background technology
Dabie Mountain Chinese ilex (Ilex dabieshanensis K Yao.et M.P.Deng.) belongs to Aquifoliaceae (Aquifoliaceae) Ilex (Ilex) plant.Evergreen dungarunga, high about 5 meters, complete stool does not have hair, and the bark canescence is level and smooth; The sprig kermesinus, the tool corner angle, the leaf keratin, ovum shape Long Circle, avette or oval, it is pointed that the anxious point of tip is thorn, the base portion subcircular, the edge is warp slightly, tool 4-8 is to ratchet, above bottle green, tool gloss, the florescence 3-4 month, the fruit phase 10-11 month, the nearly spheroidal of drupe is extremely oval, cerise when ripe.Anti-pruning; Happiness light anti-half is cloudy, cold-resistant drought-enduring; Soil is less demanding, acid and contain riotous growth in the many soil of humus, requires fertile, moistening and draining is good; Happiness warm and moist weather, year-round average temperature is advisable for 16 ℃, and the ability maximum temperature is 39 ℃, and is more cold-resistant, and-8 ℃ are not subjected to freeze injury yet, and the florescence is met the growth that low temperature can influence bloom pollination and flower.
Dabie Mountain Chinese ilex 1# is the improved seeds that the Jiangsu Province Chinese Academy of Sciences independently selects, and these seeds are suitable for the flower garden greening, view and admire, evergreen all the year round, the leaf look verdant, and fruit is red gorgeous, heat-resisting, drought-enduring, be suitable for garden, lawn isolated planting, hedgerow planting, the greening of highway both sides, be all to be subjected to both at home and abroad one of good evergreen broad-leaved seeds that people like deeply, also being the good material of making potted landscape, is one of new, excellent afforestation seeds of developing in recent years, has broad application prospects.
At present this kind seedling quantity is considerably less at home, and correlative study shows that the seminal propagation of Dabie Mountain Chinese ilex 1# and cottage propagation also break through, so Dabie Mountain Chinese ilex 1# resource lacks very much, and uses and also be " have city out of stock ".
The woody plant tissure culture studies is started in early 1950s, just obtains bigger development up to the later stage seventies.At present, incomplete statistics has more than 300 kind of woody plant of genus more than 90 to obtain regeneration plant through tissue culture.And about the research of Holly tissue culture, report seldom both at home and abroad.Nearly 5 years, rarely seen have report that Chinese ilex (Ilex chinensis Sims) (is seen Wenzhou agricultural science and technology, 2007 the 4th phases, tissue culture of Chinese ilex and quick propagating technology) and white holly [Ilex verticillata (Linn.) A.Gray] (see Plant Physiology Communications, 2007 the 2nd phases, tissue culture of white holly and breeding fast) carry out tissue culture, and do not see as yet that about the rapid propagation in vitro research of Dabie Mountain Chinese ilex 1# report is arranged.
In sum, taking tissue culture propagating Dabie Mountain Chinese ilex 1# is one of important channel that solves Dabie Mountain Chinese ilex 1# shortage of resources, and exploitation and the utilization and extention of Dabie Mountain Chinese ilex 1# all had very important meaning.
Summary of the invention
The technical problem that the present invention mainly solves is the tissue culture propagation of open Dabie Mountain Chinese ilex 1#.
Adopt following technical scheme for solving this technology:
A) explant sterilization;
B) induction of bud: the explant behind the cancellation poison is cut into the long stem section of 1.0-1.5cm, wherein each stem section is with an axillalry bud, the stem section is inoculated in the induction that carries out bud in the inducing culture, the control cultivation temperature is at 20-26 ℃, cultivate 20-26h at first in the dark, normally cultivated then 25-35 days, described normal cultivation is: light application time 10-16h/ days, intensity of illumination was 1600-2000Ix;
C) successive transfer culture and propagation: with step B) in the tender tip of gained Dabie Mountain Chinese ilex 1# cutting back insert in the subculture medium and carried out successive transfer culture 25-35 days, cultivation temperature 20-26 ℃, light application time 10-16h/ days, intensity of illumination was 1600-2000Ix;
D) inducing of root: with step C) Chinese ilex 1# test-tube plantlet in gained Dabie Mountain inserts in the root media and carried out culture of rootage 10-15 days, and cultivation temperature 20-26 ℃, light application time 10-16h/ days, intensity of illumination was 1600-2000Ix.
Explant can be selected the stem section of band axillalry bud on the annual cuttage seeding current-year branch of Dabie Mountain Chinese ilex 1#; Steps A) the explant disinfecting process in is: clean with flushing with clean water, use the washing powder rinsing then once, with vibration sterilization 15-20min on the 84 thimerosal shaking tables, behind the flowing water flushing 20-30min, on the sterile working platform with 75% ethanol disinfection 50-60s, pour 0.1% mercuric chloride sterilization 10min immediately into, with sterile water washing 4 times, 3-5min vibrates at every turn at last.
Step B) and step C) in can adopt the MS minimal medium to add three parahormones such as the hormone basic element of cell division, auximone and gibberellin, step B wherein) inducing culture that adopts in is: MS+6-BA0.5mg/L+KT0.05mg/L+NAA0.05mg/L+GA30.1mg/L, step C) in the subculture medium that adopts be: MS+6-BA0.3mg/L+IBA 0.02mg/L+GA
30.3mg/L; Step D) adoptable root media is in: 1/2MS+6-BA0.2mg/L+IBA0.4mg/L; Step B simultaneously), C), D) in also can add agar 6.5g/l in the medium that adopts, sucrose 30g/l, controlling medium pH simultaneously is 5.8-6.0.
The beneficial effect of the relative prior art of the present invention is: adopt the method for tissue culture that Dabie Mountain Chinese ilex 1# is bred, can effectively obtain a large amount of Dabie Mountain Chinese ilex 1# aseptic seedling, solve the problem of Dabie Mountain Chinese ilex 1# shortage of resources; By to the choosing of explant, inducing culture, subculture medium and root media, can make that germination rate reaches 70-90%, the rate of increase reaches 5-8 doubly, and rooting rate reaches 95-100%, can obtain Dabie Mountain Chinese ilex 1# aseptic seedling efficiently.
Embodiment
Embodiment one
1, chooses the healthy and strong annual cuttage seeding of Dabie Mountain Chinese ilex 1# of 50 strains.Clip is given birth to tender taper branch then, removes blade, is explant with the stem section, and every stem section is with an axillalry bud, is about 2cm.Explant is clean with flushing with clean water, use the washing powder rinsing then once, with vibration sterilization 15min on the 84 thimerosal shaking tables.Behind the flowing water flushing 20min, with 75% ethanol disinfection 50s, pour 0.1% mercuric chloride sterilization 10min immediately on the sterile working platform, with sterile water washing 4 times, 3min vibrates at every turn at last.0.5cm is excised at the explant two ends that sterilization is crossed respectively, to remove injured part.Insert inducing culture MS+6-BA0.5mg/L+KT0.05mg/L+NAA0.05mg/L+GA
30.1mg/L in.Every bottle graft 3 strains insert 50 bottles altogether.24 ℃ of temperature, light application time 10h/ days, under the condition of light intensity 2000lx, cultivate.
2, after explant inserted medium 7d, axillalry bud began to sprout.During to 30d, pollute 10 strains, germination rate reaches 75%, obtains not first for aseptic seedling with root of 105 strains altogether.The about 4cm of aseptic seedling plant height has 3 to 6 of the axillalry buds of having sprouted on every strain stem.Blade is removed, and trunk is cut to stem with bud, is with an axillalry bud for every section, altogether 420 sections.Insert shoot proliferation medium MS+6-BA0.3mg/L+IBA 0.02mg/L+GA
30.3mg/L in, condition of culture is the same.After cultivating 15d, axillalry bud begins to go out successively the tip, and behind the 30d, the rate of increase reaches 8.6 times, on average contains 8.6 axillalry buds on promptly every strain subculture seedling, obtains 3612 of axillalry buds altogether.
3, the subculture aseptic seedling that the 1st shoot proliferation obtained is cut to the stem section with 2 or 3 axillalry buds, obtains 1204 sections of stem sections.Carry out subculture the 2nd time, axillalry bud begins the tip behind the 10d, is on average had aseptic seedling 1150 strains (all the other 20 strains are polluted, and 34 strains are not sprouted) of 8 axillalry buds behind the 30d.Repeat subculture the 3rd time, obtain 3070 sections of stem sections, obtain on average to have about 3000 strains of subculture aseptic seedling (all the other 20 strains are polluted, and 50 strains are not sprouted) of 6 axillalry buds behind the 30d.
4, will be cut into segment with 2 or 3 axillalry buds through the aseptic seedling of 3 subcultures, and insert strong plantlets and rootage medium 1/2MS+6-BA0.2mg/L+IBA0.4mg/L, and insert 6000 strains altogether, condition of culture is the same.After cultivating 10d, begin to take root, rooting rate reaches 100% behind the 15d, so far obtains 6000 strains of band root aseptic seedling.
Also add agar 6.5g/l in the above medium, sucrose 30g/l, controlling medium pH simultaneously is 5.8-6.0.
Embodiment two
1, chooses the healthy and strong annual cuttage seeding of Dabie Mountain Chinese ilex 1# of 30 strains.The clip acrial part is removed blade, is explant with the stem section, and every stem section is with an axillalry bud, is about 2cm.Explant is clean with flushing with clean water, use the washing powder rinsing then once, with vibration sterilization 18min on the 84 thimerosal shaking tables.Behind the flowing water flushing 25min, with 75% ethanol disinfection 55s, pour 0.1% mercuric chloride sterilization 10min immediately on the sterile working platform, with sterile water washing 4 times, 4min vibrates at every turn at last.0.5cm is excised at the explant two ends that sterilization is crossed respectively, to remove injured part.Insert inducing culture MS+6-BA0.5mg/L+KT0.05mg/L+NAA0.05mg/L+GA
30.1mg/L in.Every bottle graft 3 strains insert 50 bottles altogether.25 ℃ of temperature, light application time 16h/ days, under the condition of light intensity 1600lx, cultivate.
2, after explant inserted medium 9d, axillalry bud began to sprout.During to 35d, pollute 20 strains, germination rate reaches 80%, obtains not first for aseptic seedling with root of 104 strains altogether.The average plant height 4.5cm of aseptic seedling has 3 to 6 of the axillalry buds of having sprouted on every strain stem.Blade is removed, and trunk is cut to stem with bud, is with an axillalry bud for every section, obtains 420 sections of stem sections altogether.Insert shoot proliferation medium MS+6-BA0.3mg/L+IBA 0.02mg/L+GA
30.3mg/L in, condition of culture is the same.After cultivating 10d, axillalry bud begins to go out successively the tip, and behind the 30d, the rate of increase reaches 7.2 times, on average contains 7.2 axillalry buds on promptly every strain subculture seedling, altogether 3024 of axillalry buds.
3, the subculture aseptic seedling that the 1st shoot proliferation obtained is cut to the stem section with 2 or 3 axillalry buds, obtains 1008 sections of stem sections.Carry out subculture the 2nd time, axillalry bud begins the tip behind the 10d, is on average had aseptic seedling 968 strains (all the other 10 strains are polluted, and 30 strains are not sprouted) of 8 axillalry buds behind the 30d.Repeat subculture the 3rd time, obtain 2581 sections of stem sections, obtain on average to have about 2500 strains of subculture aseptic seedling (all the other 11 strains are polluted, and 71 strains are not sprouted) of 6 axillalry buds behind the 30d.
4, will be cut into segment with 2 or 3 axillalry buds through the aseptic seedling of 3 subcultures, and insert strong plantlets and rootage medium 1/2MS+6-BA0.2mg/L+IBA0.4mg/L, and insert 5000 strains altogether, condition of culture is the same.Begin to take root after cultivating 10d, rooting rate reaches 97% behind the 15d.So far obtain to contain 4850 strains of root aseptic seedling.
Also add agar 6.5g/l in the above medium, sucrose 30g/l, controlling medium pH simultaneously is 5.8-6.0.
Embodiment three
1, chooses the healthy and strong annual cuttage seeding of Dabie Mountain Chinese ilex 1# of 40 strains.The clip acrial part is removed blade, is explant with the stem section, and every stem section is with an axillalry bud, is about 2cm.Explant is clean with flushing with clean water, use the washing powder rinsing then once, with vibration sterilization 20min on the 84 thimerosal shaking tables.Behind the flowing water flushing 30min, with 75% ethanol disinfection 60s, pour 0.1% mercuric chloride sterilization 10min immediately on the sterile working platform, with sterile water washing 4 times, 5min vibrates at every turn at last.0.5cm is excised at the explant two ends that sterilization is crossed respectively, to remove injured part.Insert inducing culture MS+6-BA0.5mg/L+KT0.05mg/L+NAA0.05mg/L+GA
30.1mg/L in.Every bottle graft 3 strains insert 50 bottles altogether.26 ℃ of temperature, light application time 16h/ days, under the condition of light intensity 1600lx, cultivate.
2, after explant inserted medium 10d, axillalry bud began to sprout.During to 35d, pollute 10 strains, germination rate reaches 87.10%, obtains not first for aseptic seedling with root of 122 strains altogether.The average plant height 4.5cm of aseptic seedling has 3 to 6 of the axillalry buds of having sprouted on every strain stem.Blade is removed, and trunk is cut to stem with bud, is with an axillalry bud for every section, altogether 488 sections of stem sections.Insert shoot proliferation medium MS+6-BA0.3mg/L+IBA 0.02mg/L+GA
30.3mg/L in, condition of culture is the same.After cultivating 13d, axillalry bud begins to go out successively the tip, and the rate of increase reaches 5.3 times behind the 30d, and promptly every strain subculture seedling on average contains 5.3 axillalry buds, altogether 2585 of axillalry buds.
3, the subculture aseptic seedling that the 1st shoot proliferation obtained is cut to the stem section with 2 or 3 axillalry buds, obtains 861 sections of stem sections.Carry out subculture the 2nd time, axillalry bud begins the tip behind the 5d, is on average had aseptic seedling 800 strains (all the other 61 strains are not sprouted) of 8 axillalry buds behind the 30d.Repeat subculture the 3rd time, obtain 2133 sections of stem sections, obtain on average to have about 2100 strains of subculture aseptic seedling (all the other 33 strains are not sprouted) of 6 axillalry buds behind the 30d.
4, will be cut into segment with 3 axillalry buds through the aseptic seedling of 3 subcultures, and insert strong plantlets and rootage medium 1/2MS+6-BA0.2mg/L+IBA0.4mg/L, and insert 4200 strains altogether, condition of culture is the same.Begin to take root after cultivating 10d, rooting rate reaches 95% behind the 15d.So far obtain to contain 3990 strains of root aseptic seedling.
Also add agar 6.5g/l in the above medium, sucrose 30g/l, controlling medium pH simultaneously is 5.8-6.0.
Claims (7)
1. Dabie Mountain Chinese ilex 1# tissue culture propagation is characterized in that, may further comprise the steps:
A) explant sterilization;
B) induction of bud: the explant behind the cancellation poison is cut into the long stem section of 1.0-1.5cm, wherein each stem section is with an axillalry bud, the stem section is inoculated in the induction that carries out bud in the inducing culture, the control cultivation temperature is at 20-26 ℃, cultivate 20-26h at first in the dark, normally cultivated then 25-35 days, described normal cultivation is: light application time 10-16h/ days, intensity of illumination was 1600-2000Ix;
C) successive transfer culture and propagation: with step B) in the tender tip of gained Dabie Mountain Chinese ilex 1# cutting back insert in the subculture medium and carried out successive transfer culture 25-35 days, cultivation temperature 20-26 ℃, light application time 10-16h/ days, intensity of illumination was 1600-2000Ix;
D) inducing of root: with step C) Chinese ilex 1# test-tube plantlet in gained Dabie Mountain inserts in the root media and carried out culture of rootage 10-15 days, and cultivation temperature 20-26 ℃, light application time 10-16h/ days, intensity of illumination was 1600-2000Ix.
2. Dabie Mountain according to claim 1 Chinese ilex 1# tissue culture propagation is characterized in that steps A) in explant choose the annual cuttage seeding of Dabie Mountain Chinese ilex 1#.
3. Dabie Mountain according to claim 1 and 2 Chinese ilex 1# tissue culture propagation, it is characterized in that, steps A) the explant disinfecting process in is: clean with flushing with clean water, use the washing powder rinsing then once, with vibration sterilization 15-20min on the 84 thimerosal shaking tables, behind the flowing water flushing 20-30min, on the sterile working platform with 75% ethanol disinfection 50-60s, pour 0.1% mercuric chloride sterilization 10min immediately into, with sterile water washing 4 times, 3-5min vibrates at every turn at last.
4. Dabie Mountain according to claim 1 Chinese ilex 1# tissue culture propagation is characterized in that step B) in the inducing culture that adopts be: MS+6-BA0.5mg/L+KT0.05mg/L+NAA0.05mg/L+GA
30.1mg/L.
5. Dabie Mountain according to claim 1 Chinese ilex 1# tissue culture propagation is characterized in that step C) in the subculture medium that adopts be: MS+6-BA0.3mg/L+IBA 0.02mg/L+GA
30.3mg/L.
6. Dabie Mountain according to claim 1 Chinese ilex 1# tissue culture propagation is characterized in that step D) in the root media that adopts be: 1/2MS+6-BA0.2mg/L+IBA0.4mg/L.
7. according to any one described Dabie Mountain Chinese ilex 1# tissue culture propagation in the claim 4 to 6, it is characterized in that also contain agar 6.5g/l in the described medium, sucrose 30g/l, medium pH are 5.8-6.0.
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| CN103563745A (en) * | 2013-10-12 | 2014-02-12 | 杭州市园林绿化股份有限公司 | Tissue culture method of ilex verticillata |
| CN104255491A (en) * | 2014-09-19 | 2015-01-07 | 江苏农林职业技术学院 | Ilex crenata golden gem tissue culture breeding method |
| CN107173231A (en) * | 2017-07-03 | 2017-09-19 | 甘肃省科学院生物研究所 | A kind of method of the Mongolian Ammopiptanthus mongolicus rapid propagation in vitro of rare or endangered species |
| CN108029559A (en) * | 2017-12-29 | 2018-05-15 | 南京林业大学 | A kind of method of quickly breeding bearberry tissue-cultured seedling |
| CN112616659A (en) * | 2020-12-14 | 2021-04-09 | 江苏省中国科学院植物研究所 | Method for somatic embryogenesis and plant regeneration of ilex davidii |
| CN115918532A (en) * | 2022-10-14 | 2023-04-07 | 江苏省中国科学院植物研究所 | Rapid propagation method of ilex davidii |
| CN115956503A (en) * | 2022-11-28 | 2023-04-14 | 江苏省中国科学院植物研究所 | Method for establishing embryonic suspension line and regenerating plant of ilex davidii |
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| CN103563745A (en) * | 2013-10-12 | 2014-02-12 | 杭州市园林绿化股份有限公司 | Tissue culture method of ilex verticillata |
| CN103563745B (en) * | 2013-10-12 | 2015-10-21 | 杭州市园林绿化股份有限公司 | A kind of method for tissue culture of ilex verticillata |
| CN104255491A (en) * | 2014-09-19 | 2015-01-07 | 江苏农林职业技术学院 | Ilex crenata golden gem tissue culture breeding method |
| CN104255491B (en) * | 2014-09-19 | 2016-03-30 | 江苏农林职业技术学院 | A kind of gold leaf tortoise plastron Chinese ilex tissue culture propagation method |
| CN107173231A (en) * | 2017-07-03 | 2017-09-19 | 甘肃省科学院生物研究所 | A kind of method of the Mongolian Ammopiptanthus mongolicus rapid propagation in vitro of rare or endangered species |
| CN108029559A (en) * | 2017-12-29 | 2018-05-15 | 南京林业大学 | A kind of method of quickly breeding bearberry tissue-cultured seedling |
| CN112616659A (en) * | 2020-12-14 | 2021-04-09 | 江苏省中国科学院植物研究所 | Method for somatic embryogenesis and plant regeneration of ilex davidii |
| CN115918532A (en) * | 2022-10-14 | 2023-04-07 | 江苏省中国科学院植物研究所 | Rapid propagation method of ilex davidii |
| CN115956503A (en) * | 2022-11-28 | 2023-04-14 | 江苏省中国科学院植物研究所 | Method for establishing embryonic suspension line and regenerating plant of ilex davidii |
| CN115956503B (en) * | 2022-11-28 | 2023-10-27 | 江苏省中国科学院植物研究所 | Method for establishing embryogenic suspension system of ilex latifolia and regenerating plants |
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